Zoltán Kis

• Ph.D. M.Sc. B.Eng.
• Senior Lecturer at The University of Sheffield

mRNA technology has significantly changed the timeline for developing and delivering a new vaccine from years to months, as demonstrated by the development and approval of two highly efficacious vaccines...

Decades of research on bacteriophage-derived RNA polymerases (RNAPs) were vital for synthesizing mRNA using the in vitro transcription (IVT) reaction for vaccines during the COVID-19 pandemic. The future success of mRNA-based products relies on the efficiency of its manufacturing process. mRNA manufacturing is a platform technology that complements...

Following the recent COVID-19 pandemic, mRNA manufacturing processes are being actively developed and optimized to produce the next generation of mRNA vaccines and therapeutics. Herein, the performance of the tangential flow filtration (TFF) was evaluated for high-recovery, and high-purity separation of mRNA from unreacted nucleoside triphosphates...

The in vitro transcription (IVT) process is a critical step in RNA production. To ensure the efficiency of RNA manufacturing, it is essential to optimize and identify its key influencing factors. In this study, multiple Gaussian Process (GP) models are used to perform efficient optimization and global sensitivity analysis (GSA). Firstly, multiple G...

Oligo-deoxythymidine (oligo-dT) ligand-based affinity chromatography is a robust method for purifying mRNA drug substances within the manufacturing process of mRNA-based products, including vaccines and therapeutics. However, the conventional batch mode of operation for oligo-dT chromatography has certain drawbacks that reduce the productivity of t...

In the dynamic landscape of industrial evolution, Industry 4.0 (I4.0) presents opportunities to revolutionise products, processes, and production. It is now clear that enabling technologies of this paradigm, such as the industrial internet of things (IIoT), artificial intelligence (AI), and Digital Twins (DTs), have reached an adequate level of tec...

Quality by digital design (QbDD) utilizes data-driven, mechanistic, or hybrid models to define and optimize a manufacturing design space. It improves upon the QbD approach used extensively in the pharmaceutical industry. The computational models developed in this approach identify and quantify the relationship between the product’s critical quality...

Quality by digital design (QbDD) is an improvement over the quality by design (QbD) paradigm. Here, computational models are used to characterize, monitor, control, and improve manufacturing processes. A few ways in which these computational models are implemented include digital twins (DT) and soft sensors. These models can be used for a wide rang...

In recent years, Generalized Modules for Membrane Antigens (GMMA) have received increased attention as an innovative vaccine platform against bacterial pathogens, particularly attractive for low- and middle-income countries because of manufacturing simplicity. The assessment of critical quality attributes (CQAs), product-process interactions, ident...

mRNA technology has recently demonstrated the ability to significantly change the timeline for developing and delivering a new vaccine from years to months. The potential of mRNA technology for rapid vaccine development has recently been highlighted by the successful development and approval of two mRNA vaccines for COVID-19. Importantly, this RNA-...

Synthetic mRNA is currently produced in standardized in vitro transcription systems. However, this one‐size‐fits‐all approach has associated drawbacks in supply chain shortages, high reagent costs, complex product‐related impurity profiles, and limited design options for molecule‐specific optimization of product yield and quality. Herein, we descri...

The simultaneous administration of SARS-CoV-2 and influenza vaccines is being carried out for the first time in the UK and around the globe in order to mitigate the health, economic, and societal impacts of these respiratory tract diseases. However, a systematic approach for planning the vaccine distribution and administration aspects of the vaccin...

The need to increase the COVID-19 vaccine manufacturing capacity at low to middle-income countries (LMIC) led to a growing focus on Novavax (NVX-CoV2373), a thermostable protein subunit vaccine manufactured using a baculovirus and insect cell system (BICS) platform. This study aimed to conduct a techno-economic analysis to assess the BICS platform...

Rapid global COVID-19 pandemic response by mass vaccination is currently limited by the rate of vaccine manufacturing. This study presents a techno-economic feasibility assessment and comparison of three vaccine production platform technologies deployed during the COVID-19 pandemic: (1) adenovirus-vectored (AVV) vaccines, (2) messenger RNA (mRNA) v...

Introduction Prior to the emergence of SARS-CoV-2, the potential use of mRNA vaccines for a rapid pandemic response had been well described in the scientific literature, however during the SARS-CoV-2 outbreak we witnessed the large-scale deployment of the platform in a real pandemic setting. Of the three RNA platforms evaluated in clinical trials,...

The COVID-19 crisis has highlighted the critical role of vaccine manufacturing in managing infectious disease outbreaks and pandemics. Enhancing RNA manufacturing capability, distribution and flexibility will now be central in future epidemic preparedness and emergency response strategies. This insight showcases the adaptation of Quality by Design...

Group A Streptococcus (GAS) causes about 500,000 annual deaths globally, and no vaccines are currently available. The Group A Carbohydrate (GAC), conserved across all GAS serotypes, conjugated to an appropriate carrier protein, represents a promising vaccine candidate. Here, we explored the possibility to use Generalized Modules for Membrane Antige...

Lipid nanoparticles (LNPs) are the leading technology for RNA delivery, given the success of the Pfizer/BioNTech and Moderna COVID-19 mRNA (mRNA) vaccines, and small interfering RNA (siRNA) therapies (patisiran). However, optimization of LNP process parameters and compositions for larger RNA payloads such as self-amplifying RNA (saRNA), which can h...

RNA-based products have emerged as one of the most promising and strategic technologies for global vaccination, infectious disease control, and future therapy development. The assessment of critical quality attributes (CQAs), product-process interactions, relevant process analytical technologies, and process modeling capabilities can feed into a ro...

The transition to energy systems with a high share of renewable energy depends on the availability of technologies that can connect the physical distances or bridge the time differences between the energy supply and demand points. This study focuses on energy storage technologies due to their expected role in liberating the energy sector from fossi...

The vaccine distribution chains in several low- and middle-income countries are not adequate to facilitate the rapid delivery of high volumes of thermosensitive COVID-19 mRNA vaccines at the required low and ultra-low temperatures. COVID-19 mRNA vaccines are currently distributed along with temperature monitoring devices to track and identify devia...

Vaccine production platform technologies have played a crucial role in rapidly developing and manufacturing vaccines during the COVID-19 pandemic. The role of disease agnostic platform technologies, such as the adenovirus-vectored (AVV), messenger RNA (mRNA), and the newer self-amplifying RNA (saRNA) vaccine platforms is expected to further increas...

This work develops a multi-product MILP vaccine supply chain model that supports planning, distribution, and administration of viral vectors and RNA-based vaccines. The capability of the proposed vaccine supply chain model is illustrated using a real-world case study on vaccination against SARS-CoV-2 in the UK that concerns both viral vectors (e.g....

Vaccination plays a key role in reducing morbidity and mortality caused by infectious diseases, including the recent COVID-19 pandemic. However, a comprehensive approach that allows the planning of vaccination campaigns and the estimation of the resources required to deliver and administer COVID-19 vaccines is lacking. This work implements a new fr...

Rapid global COVID-19 pandemic response by mass vaccination is currently limited by the rate of vaccine manufacturing. This study presents a techno-economic feasibility assessment and comparison of three vaccine production platform technologies deployed during the COVID-19 pandemic: (1) adenovirus-vectored (AVV) vaccines, (2) messenger RNA (mRNA) v...

Rapid-response vaccine production platform technologies, including RNA vaccines, are being developed to combat viral epidemics and pandemics. A key enabler of rapid response is having quality-oriented disease-agnostic manufacturing protocols ready ahead of outbreaks. We are the first to apply the Quality by Design (QbD) framework to enhance rapid-r...

The authors wish to make the following corrections to this paper [...]

Pandemics such as the current COVID-19 outbreak pose tremendous healthcare and economic challenges. Vaccines hold promise for controlling pandemics; however, substantial challenges come with pandemic-response vaccine development, manufacturing, distribution, and administration. To address those now, many companies are using rapid-response vaccine-p...

To overcome pandemics, such as COVID-19, vaccines are urgently needed at very high volumes. Here we assess the techno-economic feasibility of producing RNA vaccines for the demand associated with a global vaccination campaign. Production process performance is assessed for three messenger RNA (mRNA) and one self-amplifying RNA (saRNA) vaccines, all...

Supplementary Informtion (SI) document for "Resources, production scales and time required for producing RNA vaccines for the global pandemic demand"

Overcoming pandemics, such as the current Covid‐19 outbreak, requires the manufacture of several billion doses of vaccines within months. This is an extremely challenging task given the constraints in small‐scale manufacturing for clinical trials, clinical testing timelines involving multiple phases and large‐scale drug substance and drug product m...

Well-established and newly developed genome technologies are revolutionising the field of biomedicine, by providing genomic data and genetic engineered structures that support investigating individual propensity for developing certain diseases, on one hand, and by predicting individual responses to the environmental stimulus due to gene common vari...

Well-established and newly developed genome technologies are revolutionising the field of biomedicine, by providing genomic data and genetic engineered structures that support investigating individual propensity for developing certain diseases, on one hand, and by predicting individual responses to the environmental stimulus due to gene common vari...

Vaccine manufacture currently follows a centralized approach dominated by large‐scale, nonflexible, and product‐specific facilities, which require high investment costs. Emerging vaccine platform technologies, such as RNA vaccines, outer membrane vesicle vaccines with genetically customizable membrane antigens (customOMV), virus‐like particle vacci...

To stop the spread of future epidemics and meet infant vaccination demands in low- and middle-income countries, flexible, rapid and low-cost vaccine development and manufacturing technologies are required. Vaccine development platform technologies that can produce a wide range of vaccines are emerging, including: a) humanized, high-yield yeast reco...

Improvements in water, sanitation and hygiene (WASH) service provision are hampered by limited open data availability. This paper presents a data integration framework, collects the data and develops a material flow model, which aids data-based policy and infrastructure development for the WASH sector. This model provides a robust quantitative mapp...

About 70% of all Ewing sarcoma (EWS) patients are diagnosed under the age of 20 years. Over the last decades little progress has been made towards finding effective treatment approaches for primarily metastasized or refractory Ewing sarcoma in young patients. Here, in the context of the search for novel therapeutic options, the potential of oncolyt...

Osteosarcoma is the most frequent malignant disease of the bone. On the basis of early clinical experience in the 1960s with H-1 protoparvovirus (H-1PV) in osteosarcoma patients, this effective oncolytic virus was selected for systematic preclinical testing on various osteosarcoma cell cultures. A panel of five human osteosarcoma cell lines (CAL 72...

The majority of (mammalian) cells in our body are sensitive to mechanical forces, but little work has been done to develop assays to monitor mechanosensor activity. Furthermore, it is currently impossible to use mechanosensor activity to drive gene expression. To address these needs, we developed the first mammalian mechanosensitive synthetic gene...

The discovery of the human genome has unveiled new fields of genomics, transcriptomics, and proteomics, which has produced paradigm shifts on how to study disease mechanisms, wherein a current central focus is the understanding of how gene signatures and gene networks interact within cells. These gene function studies require manipulating genes eit...

In this review, we discuss new emerging medical applications of the rapidly evolving field of mammalian synthetic biology. We start with simple mammalian synthetic biological components and move towards more complex and therapy-oriented gene circuits. A comprehensive list of ON-OFF switches, categorized into transcriptional, post-transcriptional, t...

Employing synthetic biology and shear stress sensing to tackle atherosclerosis Oliver Fleck§, Zoltán Kis§, Asma Zafar§¥, Zhangxing Lai§£, Takayuki Homma§ and Rob Krams§ §Department of Bioengineering, Imperial College London, UK ¥Government College University, Lahore, Pakistan £Department of Mechanical Engineering, National University of Singapore,...

The ability of cells to receive and respond to chemical and physical signals from beyond the plasma membrane is fundamental to life. Sensing and processing of stimuli (such as hormones, mechanical factors, light, heat, neurotransmitters, nutrients, odorants, etc.) is mediated by signal-transduction pathways, molecular circuits that detect, amplify,...

Atherosclerosis is intimately coupled to blood flow by the presence of predilection sites. The coupling is through mechanotransduction of endothelial cells and approximately 2000 gene are associated with this process. This paper describes a new platform to study and identify new signalling pathways in endothelial cells covering an atherosclerotic p...

EPR spectroscopy is employed to demonstrate chemical production of formally Fe(i) and Fe(0) states of phthalocyanines in water at room temperature, and physiologically-relevant pH.

Thiourea dioxide, (NH2)2CSO2 (TDO), is known to decompose in alkaline media to yield the strong reducing agent sulfoxylate, SO2H−, with interesting applications in (bio)catalysis. The electronic structure of TDO has been debated recently, with two different descriptions put forth based on crystal structures and electronic structure calculations, al...

Florina Deac, Nicoleta Cotolan, Zoltán Kis, Radu Silaghi-Dumitrescu

Deoxy hemoglobin is well-known to be pentacoordinated, with axial ligation from the so-called proximal histidine. The reducing agent, dithionite, is commonly employed to generate the deoxy from in hemoglobin and other proteins. Here, we report that at room temperature in strongly alkaline medium hemoglobin but not myoglobin can be converted by redu...

Recent experimental data from us and others indicate formation of Fe(0) complexes of biologically relevant systems, which may be involved in carbon dioxide reduction to CO. Here, density functional theory results are presented in heme Fe(0) models (relevant for proteins such as hemoglobin and peroxidases), including adducts with carbonate/CO(2) and...

Recent results from our group on super-reduced (Fe(0), Fe(I)) as well as on high-valent Fe(IV) are reviewed. These include density functional calculations, UV-vis, NMR and EPR spectra, stopped-flow kinetics, and enzyme kinetics. The targets include globins (with relevance for blood substitute analysis, among others), cytochrome c (with possible rel...

Sulfoxylate SO2H−(SO22−), a strong reducing agent readily produced by hydrolysis of thiourea dioxide, reacts with ferric myoglobin (Mb) to reversibly produce Fe(II)-Mb, starting from either aerobic or anaerobic conditions. Exposure of Fe(II)-Mb to excess sulfoxylate further produces Fe(II)-CO-Mb. Fe(II)-Mb can be regenerated by reoxidation with fer...